Characterization of EGFR-reprogrammable temozolomide-resistant cells in a model of glioblastoma.
Characterization of EGFR-reprogrammable temozolomide-resistant cells in a model of glioblastoma.
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胶质母细胞瘤模型中 EGFR 可重编程替莫唑胺耐药细胞的表征
DOI：
10.1038/s41420-022-01230-y
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发表时间：
2022-10-31
影响因子：
7
通讯作者：
Zou J
中科院分区：
文献类型：
--
作者：
Gong L;Yin Y;Chen C;Wan Q;Xia D;Wang M;Pu Z;Zhang B;Zou J
替莫唑胺（TMZ）耐药是胶质母细胞瘤（GBM）的主要临床挑战。O 6-甲基鸟嘌呤-DNA甲基转移酶（MGMT）介导的DNA损伤修复是TMZ耐药的重要机制。然而，MGMT缺失的GBM患者仍然对TMZ耐药，并且耐药演变的过程在很大程度上是未知的。在这里，我们开发了一种获得性TMZ耐药异种移植模型，使用MGMT-高甲基化U87细胞的连续植入，允许提取稳定的TMZ耐药（TMZ-R）肿瘤和原代细胞。衍生的肿瘤和细胞在体外和体内均表现出稳定的多药耐药性。功能实验以及单细胞RNA测序（scRNA-seq）表明，TMZ治疗诱导了细胞异质性，包括TMZ-R肿瘤中的静止癌干细胞（CSC）。其中一个亚组被内斯+/SOX 2 +/CADM 1+标记，并表现出显著的耐药性优势。进一步的研究表明，表皮生长因子受体（EGFR）的缺陷和下游信号的减少可能会赋予这个三重阳性的CSC亚群的静止表型和化疗耐药性。连续EGF治疗提高TMZ-R细胞在体外和体内的化疗敏感性，机械逆转细胞周期停滞和减少药物摄取。此外，TMZ-R肿瘤的EGF治疗有利地使联合治疗中对TMZ的反应正常化。在这里，我们描述了一个独特的亚组的CSC在MGMT无效的实验性胶质母细胞瘤，确定EGF + TMZ治疗作为一个潜在的策略，以克服细胞静止和TMZ耐药，可能赋予缺陷的EGFR信号。
Temozolomide (TMZ) resistance is a major clinical challenge for glioblastoma (GBM). O6-methylguanine-DNA methyltransferase (MGMT) mediated DNA damage repair is a key mechanism for TMZ resistance. However, MGMT-null GBM patients remain resistant to TMZ, and the process for resistance evolution is largely unknown. Here, we developed an acquired TMZ resistant xenograft model using serial implantation of MGMT-hypermethylated U87 cells, allowing the extraction of stable, TMZ resistant (TMZ-R) tumors and primary cells. The derived tumors and cells exhibited stable multidrug resistance both in vitro and in vivo. Functional experiments, as well as single-cell RNA sequencing (scRNA-seq), indicated that TMZ treatment induced cellular heterogeneity including quiescent cancer stem cells (CSCs) in TMZ-R tumors. A subset of these were labeled by NES+/SOX2+/CADM1+ and demonstrated significant advantages for drug resistance. Further study revealed that Epidermal Growth Factor Receptor (EGFR) deficiency and diminished downstream signaling may confer this triple positive CSCs subgroup’s quiescent phenotypes and chemoresistance. Continuous EGF treatment improved the chemosensitivity of TMZ-R cells both in vitro and in vivo, mechanically reversing cell cycle arrest and reduced drug uptake. Further, EGF treatment of TMZ-R tumors favorably normalized the response to TMZ in combination therapy. Here, we characterize a unique subgroup of CSCs in MGMT-null experimental glioblastoma, identifying EGF + TMZ therapy as a potential strategy to overcome cellular quiescence and TMZ resistance, likely endowed by deficient EGFR signaling.
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影响因子：
3.4
作者：
Grogan, Patrick T.;Sarkaria, Jann N.;Timmermann, Barbara N.;Cohen, Mark S.
通讯作者：
Cohen, Mark S.
影响因子：
5.6
作者：
Alimbetov D;Askarova S;Umbayev B;Davis T;Kipling D
通讯作者：
Kipling D
影响因子：
64.5
作者：
Asp, Michaela;Giacomello, Stefania;Lundeberg, Joakim
通讯作者：
Lundeberg, Joakim
影响因子：
4.6
作者：
Areeb Z;Stuart SF;West AJ;Gomez J;Nguyen HPT;Paradiso L;Zulkifli A;Jones J;Kaye AH;Morokoff AP;Luwor RB
通讯作者：
Luwor RB
影响因子：
158.5
作者：
Hegi, ME;Diserens, A;Stupp, R
通讯作者：
Stupp, R